Velocity, lifetime, and Z ring morphology measurements for cells
missing each synthetic lethal combination of ZBPs. All synthetic lethal
combinations were investigated by a combination of knockouts (indicated by
Δ) and depletions (indicated by ↓); depletions were performed
by expressing the gene under an inducible promoter until the start of the
experiment, then withdrawing the inducer for 7 hours. a
Velocity (left) and lifetime (right) of
cells missing synthetically lethal combinations of ZBPs are unchanged from
control. For velocity measurements, FtsZ-mNeonGreen was induced with 20
μM IPTG for 2 hours, imaged by TIRFM, and then analysed from
kymographs. For lifetime measurements, FtsZ-HaloTag was induced with 20
μM IPTG for 2 hours and labelled with 40 pM JF549-HTL.
bc Z rings in cells missing synthetically lethal
combinations of ZBPs are wider than control cells and cells missing
individual ZBPs. Average intensity projections (b) and widths
(c) of Z rings in each condition. Z rings were visualized
using epifluorescence images of cells expressing FtsZ-mNeonGreen, induced
with 20 μM IPTG for 2 hours. Z ring projections were created by
averaging >100 Z ring images for each strain. Because ZBPs can be
removed by either knockout or depletion, for each strain we compare to the
equivalent single mutant knockouts and depletions.